ArticlePloS one2015
Efficient fdCas9 Synthetic Endonuclease with Improved Specificity for Precise Genome Engineering.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
What it found
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Who cites it
26 citing papers in PubMed, 51 citations in OpenAlex.
- CRISPR and the Future of Cardiac Disease Therapy: A New Genetic Frontier.International journal of molecular sciences · 2026Review
- Precise, specific gene editing via a compact GoCas12m-FokI chimeric nuclease.Nucleic acids research · 2026Article
- Progress and pitfalls of gene editing technology in CAR-T cell therapy: a state-of-the-art review.Frontiers in oncology · 2024Review
- Application and perspective of CRISPR/Cas9 genome editing technology in human diseases modeling and gene therapy.Frontiers in genetics · 2024Review
- CRISPR/Cas9 Landscape: Current State and Future Perspectives.International journal of molecular sciences · 2023Review
- A simple protocol to isolate a single human cell PRDX1 knockout generated by CRISPR-Cas9 system.STAR protocols · 2022Article
- Rapid Assessment of CRISPR Transfection Efficiency and Enrichment of CRISPR Induced Mutations Using a Dual-Fluorescent Stable Reporter System.Frontiers in genome editing · 2022Article
- Genome Editing and Designer Crops for the Future.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Review
- Tissue-Specific Delivery of CRISPR Therapeutics: Strategies and Mechanisms of Non-Viral Vectors.International journal of molecular sciences · 2020Review
- Efficient CRISPR/Cas9-Mediated Mutagenesis in Primary Murine T Lymphocytes.Current protocols in immunology · 2019Article
- Plant Genome Engineering for Targeted Improvement of Crop Traits.Frontiers in plant science · 2019Review
- Virus-Mediated Genome Editing in Plants Using the CRISPR/Cas9 System.Methods in molecular biology (Clifton, N.J.) · 2019Article
- Multiplexed editing of a begomovirus genome restricts escape mutant formation and disease development.PloS one · 2019Article
- NmeCas9 is an intrinsically high-fidelity genome-editing platform.Genome biology · 2018Article
- Applications of Alternative Nucleases in the Age of CRISPR/Cas9.International journal of molecular sciences · 2017Review
- Targeted insertion of an anti-CD2 monoclonal antibody transgene into the GGTA1 locus in pigs using FokI-dCas9.Scientific reports · 2017Article
- CRISPR-Cas orthologues and variants: optimizing the repertoire, specificity and delivery of genome engineering tools.Mammalian genome : official journal of the International Mammalian Genome Society · 2017Review
- Efficient CRISPR/Cas9-Mediated Genome Editing Using a Chimeric Single-Guide RNA Molecule.Frontiers in plant science · 2017Article
- Biasing genome-editing events toward precise length deletions with an RNA-guided TevCas9 dual nuclease.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
Corrections and comments
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Authors and funding
12 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The Cas9 endonuclease is used for genome editing applications in diverse eukaryotic species. A high frequency of off-target activity has been reported in many cell types, limiting its applications to genome engineering, especially in genomic medicine. Here, we generated a synthetic chimeric protein between the catalytic domain of the FokI endonuclease and the catalytically inactive Cas9 protein (fdCas9). A pair of guide RNAs (gRNAs) that bind to sense and antisense strands with a defined spacer sequence range can be used to form a catalytically active dimeric fdCas9 protein and generate double-strand breaks (DSBs) within the spacer sequence. Our data demonstrate an improved catalytic activity of the fdCas9 endonuclease, with a spacer range of 15-39 nucleotides, on surrogate reporters and genomic targets. Furthermore, we observed no detectable fdCas9 activity at known Cas9 off-target sites. Taken together, our data suggest that the fdCas9 endonuclease variant is a superior platform for genome editing applications in eukaryotic systems including mammalian cells.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.